Effects of miR-96-5p on high-glucose-induced keratinocytes and the underlying mechanism

ZHU Yan; JIANG Fangfang; XIONG Leilei

doi:10.3969/j.issn.1002-7386.2023.24.008

Abstract：Objective To investigate the effects of miR-96-5p on high-glucose-induced keratinocytes and the underlying mechanism.Methods Human keratinocytes were isolated and treated in the culture medium containing 50mmol/L glucose to simulate the hyperglycemic environment.Overexpression and knockdown of miR-96-5p were achieved by transfection of miR-96-5p mimic and inhibitor,respectively.Cells were treated with blank control,normal induction,high-glucose induction,high-glucose induction plus transfection of miR-96-5p mimic,and high-glucose induction plus transfection of miR-96-5p inhibitor.Cell migration,proliferation and invasion were detected by scratch assay,CCK-8 assay,and Transwell assay,respectively.Western blot was used to detect the expression levels of Bcl-2/adenovirus E1B 19-kDa interacting protein 3(BNIP3),focal adhesion kinase(FAK)and phosphorylated FAK(p-FAK).Results The induction of 50mmol/L glucose in keratinocytes results in decreased refraction and the presence of cellular cavity.Compared with cells treated with blank control and normal induction,the number of migratory and invasive cells,and proliferative rate at eat time point were significantly lower in those induced with high-glucose,high-glucose induction plus transfection of miR-96-5p mimic,and high-glucose induction plus transfection of miR-96-5p inhibitor(all P<0.05),which were significantly higher in cells with high-glucose induction plus transfection of miR-96-5p inhibitor than those induced with high-glucose,and high-glucose induction plus transfection of miR-96-5p mimic(all P<0.05).Compared with cells treated with blank control and normal induction,protein levels of BNIP3,FAK and p-FAK were significantly lower in those induced with high-glucose,high-glucose induction plus transfection of miR-96-5p mimic,and high-glucose induction plus transfection of miR-96-5p inhibitor(P<0.05),which were significantly higher in cells with high-glucose induction plus transfection of miR-96-5p inhibitor than those induced with high-glucose,and high-glucose induction plus transfection of miR-96-5p mimic(P<0.05).Conclusion MiR-96-5p can affect the proliferation,invasion and migration of keratinocytes by activating the BNIP3/FAK signaling pathway,and thus influencing the healing of diabetic foot wounds.